The 50 most-cited articles in Analytical Chemistry, 2021

Microplastic pollution research has suffered from inadequate data and tools for spectral (Raman and infrared) classification. Spectral matching tools often are not accurate for microplastics identification and are cost-prohibitive. Lack of accuracy stems from the diversity of microplastic pollutants, which are not represented in spectral libraries. Here, we propose a viable software solution: Open Specy. Open Specy is on the web (www.openspecy.org) and in an R package. Open Specy is free and allows users to view, process, identify, and share their spectra to a community library. Users can upload and process their spectra using smoothing (Savitzky-Golay filter) and polynomial baseline correction techniques (IModPolyFit). The processed spectrum can be downloaded to be used in other applications or identified using an onboard reference library and correlation-based matching criteria. Open Specy's data sharing and session log features ensure reproducible results. Open Specy houses a growing library of reference spectra, which increasingly represents the diversity of microplastics as a…

Traditional peroxidase-like nanozyme-based sensors suffer from self-decomposition and high toxicity of H 2 O 2, as well as the interference of color from nanozymes themselves and testing samples. In this work, we adopt nanozymes (two-dimension (2D) MnO 2 sheets, manganese dioxide nanosheets (MnNS)) with oxidase-like and peroxidase-like properties as advanced catalysts to develop a novel homogeneous electrochemical sensor for organophosphate pesticides (OPs) using dissolved O 2 as a coreactant without the interference of H 2 O 2 and color. Owing to the large surface area and unique catalytic activity of MnNS, a large amount of tetramethylbenzidine (TMB) is catalyzed oxidation, leading to a significantly declined differential pulse voltammetry (DPV) current. Obviously, MnNS display an excellent response to thiocholine, deriving from the catalyzing hydrolysis of acetylthiocholine (ATCh) by acetylcholinesterase (AChE), which switches a homogeneous electrochemical OP detection process based on the depressing AChE activity with a limit of detection (LOD) of 0.025 ng mL –1 . The…

mass-spectrum-to-compound identification. This work improves CFM-ID's ability to predict ESI-MS/MS spectra from compounds by (1) learning parameters from features based on the molecular topology, (2) adding a new approach to ring cleavage that models such cleavage as a sequence of simple chemical bond dissociations, and (3) expanding its hand-written rule-based predictor to cover more chemical classes, including acylcarnitines, acylcholines, flavonols, flavones, flavanones, and flavonoid glycosides. We demonstrate that this new version of CFM-ID (version 4.0) is significantly more accurate than previous CFM-ID versions in terms of both EI-MS/MS spectral prediction and compound identification. CFM-ID 4.0 is available at http://cfmid4.wishartlab.com/ as a web server and docker images can be downloaded at https://hub.docker.com/r/wishartlab/cfmid.

The exosome has emerged as a promising noninvasive biomarker for the early diagnosis of cancer. Therefore, it is highly desirable to develop simple, inexpensive, and user-friendly biosensors for convenient, sensitive, and quantitative exosome assay. Herein, we developed a simple and cost-efficient electrochemical biosensor by combining a metal–organic framework (MOF)-functionalized paper and a screen-printed electrode (SPE) for portable, ultrasensitive, and quantitative determination of cancer-derived exosomes. In principle, the biosensor relied on recognition of the exosome by Zr-MOFs and aptamer to initiate the hybridization chain reaction (HCR) and the formation of DNAzyme for signal amplification. Benefiting from the high signal amplification ability of HCR, the label-free paper-based biosensor is capable of ultrasensitive exosome assay with a detection limit down to 5 × 10 3 particles/mL, which is superior to that of most reported methods. Moreover, the proposed paper-based biosensor possessed the advantages of low cost, simple operation, and high sensitivity, making it…

Organophosphorus pesticides (OPs) can inhibit the activity of acetylcholinesterase (AChE) to induce neurological diseases. It is significant to exploit a rapid and sensitive strategy to monitor OPs. Here, a metal-organic framework (MOF) acted as a carrier to encapsulate AuNCs, which can limit the molecular motion of AuNCs, trigger the aggregation-induced emission (AIE) effect, and exhibit a strong fluorescence with a fluorescence lifetime and quantum yield of 6.83 μs and 4.63%, respectively. Then, the marriage of fluorescence and colorimetric signals was realized on the basis of the dual function of the enzymolysis product from AChE and choline oxidase (CHO) on AuNCs@ZIF-8. First, it can decompose ZIF-8 to weaken the restraint on AuNCs, and thus the fluorescence receded. Second, it can be used as a substrate for the peroxidase mimics of the released AuNCs to oxidize 3,3',5,5'-tetramethylbenzidine (TMB) and a visible blue appeared. Thus, on the basis of the inhibition of AChE activity by OPs, a fluorescence-colorimetric dual-signal biosensor was established. In addition, colorimetric…

This work demonstrated a pressure-based biosensor integrated with a flexible pressure sensor and an electrochromic device for visual detection. Initially, a sandwich-type immunoreaction for target carcinoembryonic antigen (CEA, as a model analyte) was carried out using the capture antibody (cAb) and platinum nanoparticles-labeled detection antibody (PtNPs-dAb) in a reaction cell. The added hydrogen peroxide (H 2 O 2 ) could be catalyzed by the PtNPs to generate oxygen (O 2 ). In a sealed chamber, the pressure increased with the overflowing O 2 . Meanwhile, a skin-inspired flexible pressure sensor with excellent sensing performance was fabricated to monitor the pressure change in real time. Thus, the electrical signal of the pressure sensor could reveal the target concentration. Moreover, a voltage-regulated electrochromic device based on polyaniline (PANI) and tungsten oxide (WO 3 ) was integrated into the platform to provide a visualized readout. According to the electrical signal of the pressure sensor, the electrochromic device would change its color from green to blue, which…

ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTNanozyme-Enabled Analytical ChemistrySirong LiSirong LiDepartment of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing National Laboratory of Microstructures, Jiangsu Key Laboratory of Artificial Functional Materials, Nanjing University, Nanjing, Jiangsu 210023, ChinaMore by Sirong LiView Biography, Yihong ZhangYihong ZhangState Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Chemistry and Biomedicine Innovation Center (ChemBIC), Nanjing University, Nanjing, Jiangsu 210023, ChinaMore by Yihong ZhangView Biography, Quan WangQuan WangDepartment of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing National Laboratory of Microstructures, Jiangsu Key Laboratory of Artificial Functional Materials, Nanjing University, Nanjing, Jiangsu 210023, ChinaMore by Quan WangView Biography, Anqi LinAnqi LinDepartment of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing National Laboratory of Microstructures, Jiangsu Key…

The application of metabolomics in translational research suffers from several technological bottlenecks, such as data reproducibility issues and the lack of standardization of sample profiling procedures. Here, we report an automated high-throughput metabolite array technology that can rapidly and quantitatively determine 324 metabolites including fatty acids, amino acids, organic acids, carbohydrates, and bile acids. Metabolite identification and quantification is achieved using the Targeted Metabolome Batch Quantification (TMBQ) software, the first cross-vendor data processing pipeline. A test of this metabolite array was performed by analyzing serum samples from patients with chronic liver disease ( N = 1234). With high detection efficiency and sensitivity in serum, urine, feces, cell lysates, and liver tissue samples and suitable for different mass spectrometry systems, this metabolite array technology holds great potential for biomarker discovery and high throughput clinical testing. Additionally, data generated from such standardized procedures can be used to generate a…

This Feature summarizes recent works in paper-based potentiometry and voltammetry in heavy metal determination. Interactions of paper substrates with heavy metals, influence on the sensing response, and modification methods applied to paper substrates to improve the performance of recently developed electrochemical sensors are discussed. Since the rekindling of interest in paper-based analytical devices, methodologies and electrode designs for heavy metal determinations are highlighted. Promising aspects of the use of these sensors for samples containing solids and the increased versatility of the use of paper in analytics offers the possibility of increased acceptance of these low-cost platforms.

The outbreak of the pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) calls for an urgent unmet need for developing a facial and cost-effective detection method. The requirement of well-trained personnel and sophisticated instrument of current primary mean (reverse transcription polymerase chain reaction, RT-PCR) may hinder the practical application worldwide. In this regard, a reverse transcription recombinase polymerase amplification (RT-RPA) coupled with CRISPR-Cas12a colorimetric assay is proposed for the SARS-CoV-2 detection. The methodology we have described herein utilizes DNA-modified gold nanoparticles (AuNPs) as a universal colorimetric readout and can specifically target ORF1ab and N regions of the SARS-CoV-2 genome. After the virus genome is amplified through RT-RPA, the resulting abundant dsDNA will bind and activate Cas12a. Under trans-cleavage degradation, the capped DNA substrate will be hydrolyzed gradually from AuNPs, demonstrating a change in the surface plasmon resonance (SPR), which can be facially monitored by UV-vis absorbance…

Sensitivity, selectivity, visible detection, and rapid response are the main concerns for an analytical method. Herein, we reported a metal–organic framework (MOF)-based ratiometric fluorescence detection strategy for hypochlorous acid (HClO). The MOF was prepared with dual ligands, 2-aminoterephthalic acid (BDC-NH 2 ) and dipicolinic acid (DPA) and Eu 3+ ions as a metal node, denoted as Eu-BDC-NH 2 /DPA. The dual-ligand strategy realized the dual emission for ratiometric sensing and visual detection, adjusted the size and morphology of MOFs to obtain a good dispersion for a rapid response, and provided an amino group for the special recognition of HClO. Thus, the MOF exhibited a dual emission derived from BDC–NH 2 and Eu 3+ ions at 433 and 621 nm, respectively, under a single excitation at 270 nm. A hydrogen bond forms between an −NH 2 group and HClO to weaken the blue fluorescence at 433 nm, while the antenna effect emission from Eu 3+ ions kept stable, so ratiometric sensing was realized with an easy-to-differentiate color change for visible detection. The ratiometric sensing…

Bacterial infection is one of the major causes of human death worldwide. To prevent bacterial infectious diseases from spreading, it is of critical importance to develop convenient, ultrasensitive, and cost-efficient methods for bacteria detection. Here, an electrochemical detector of a functional two-dimensional (2D) metal–organic framework (MOF) nanozyme was developed for the sensitive detection of pathogenic Staphylococcus aureus . A dual recognition strategy consisting of vancomycin and anti- S. aureus antibody was proposed to specifically anchor S. aureus . The 2D MOFs with excellent peroxidase-like activity can efficiently catalyze o -phenylenediamine to 2,2-diaminoazobenzene, which is an ideal electrochemical signal readout for monitoring the bacteria concentration. Under optimal conditions, the present bioassay provides a wide detection range of 10–7.5 × 10 7 colony-forming units CFU/mL with a detection limit of 6 CFU/mL, which is better than most of the previous reports. In addition, the established electrochemical sensor can selectively and accurately identify S. aureus in…

ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTWater Analysis: Emerging Contaminants and Current IssuesSusan D. Richardson*Susan D. RichardsonDepartment of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29205, United States*Email: [email protected]More by Susan D. RichardsonView Biographyhttps://orcid.org/0000-0001-6207-4513 and Thomas A. TernesThomas A. TernesFederal Institute of Hydrology, Am Mainzer Tor 1, Koblenz 56068, GermanyMore by Thomas A. TernesView Biographyhttps://orcid.org/0000-0002-2615-7925Cite this: Anal. Chem. 2022, 94, 1, 382–416Publication Date (Web):November 28, 2021Publication History Published online28 November 2021Published inissue 11 January 2022https://pubs.acs.org/doi/10.1021/acs.analchem.1c04640https://doi.org/10.1021/acs.analchem.1c04640review-articleACS PublicationsCopyright © 2021 American Chemical SocietyRequest reuse permissionsArticle Views6736Altmetric-Citations38LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all…

A noninvasive fiber material-based wearable electrochemical sensor to continuously monitor the glucose level in sweat is highly desirable for smart fabrics for personal diabetes management. To achieve it, the key challenge is to construct fibers with high stretchability and excellent electrochemical performance. Herein, a highly stretchable Ni–Co metal–organic framework/Ag/reduced graphene oxide/polyurethane (Ni–Co MOF/Ag/rGO/PU) fiber-based wearable electrochemical sensor is fabricated for monitoring the glucose level in sweat continuously with high sensitivity and accuracy. The rGO/PU fiber was simply produced by an improved wet spinning technology, and the Ni–Co MOF nanosheet was coated on its surface to prepare the Ni–Co MOF/Ag/rGO/PU (NCGP) fiber electrode. The Ni–Co MOF has a large specific surface area and high catalytic activity, which enables the fiber sensors with good electrochemical performance with a high sensitivity of 425.9 μA·mM –1 ·cm –2 and a wide linear range of 10 μM–0.66 mM. More importantly, the NCGP fiber electrode also shows extremely high stretching and…

Single-atom catalysts have attracted numerous attention due to the high utilization of metallic atoms, abundant active sites, and highly catalytic activities. Herein, a single-atom ruthenium biomimetic enzyme (Ru-Ala-C 3 N 4 ) is prepared by dispersing Ru atoms on a carbon nitride support for the simultaneous electrochemical detection of dopamine (DA) and uric acid (UA), which are coexisting important biological molecules involving in many physiological and pathological aspects. The morphology and elemental states of the single-atom Ru catalyst are studied by transmission electron microscopy, energy dispersive X-ray elemental mapping, high-angle annular dark field–scanning transmission electron microscopy, and high-resolution X-ray photoelectron spectroscopy. Results show that Ru atoms atomically disperse throughout the C 3 N 4 support by Ru–N chemical bonds. The electrochemical characterizations indicate that the Ru-Ala-C 3 N 4 biosensor can simultaneously detect the oxidation of DA and UA with a separation of peak potential of 180 mV with high sensitivity and excellent…

Loop-mediated isothermal amplification (LAMP) has been increasingly applied in nucleic acid detection for clinical diagnosis and monitoring pathogenic microorganisms due to its isothermal nature and high sensitivity. However, the false-positive signal resulting from the non-specific amplification and the complexity of primer design are still technically challenging for wide applications. In this paper, we developed the CRISPR/Cas12a-assisted sequence-specific detection of LAMP products to eliminate the effect of non-specific amplification from primer dimers and spurious amplicons. Moreover, by designing a pair of target-specific stem-loop DNA probes, we greatly simplified the primer design for LAMP. The DNA probes could be ligated to form a double-stem-loop DNA template by the detected target, which initiated LAMP reaction and achieved one-nucleotide resolution due to the highly specific ligase reaction. Using microRNAs (miRNAs) as the model targets, the CRISPR/Cas12a-assisted ligation-initiated loop-mediated isothermal amplification (CAL-LAMP) can sensitively detect as low as 0.1…

Early diagnosis of cancers relies on the sensitive detection of specific biomarkers, but most of the current testing methods are inaccessible to home healthcare due to cumbersome steps, prolonged testing time, and utilization of toxic and hazardous substances. Herein, we developed a portable self-powered photoelectrochemical (PEC) sensing platform for rapid detection of prostate-specific antigen (PSA, as a model disease-related protein) by integrating a self-powered photoelectric signal output system catalyzed with chemiluminescence-functionalized Au nanoparticles (AuNPs) and a phosphomolybdic acid (PMA)-based photochromic visualization platform. TiO 2 -g-C 3 N 4 -PMA photosensitive materials were first synthesized and functionalized on a sensor chip. The sensor consisted of filter paper modified with a photocatalytic material and a regional laser-etched FTO electrode as an alternative to a conventional PEC sensor with a glass-based electrode. The targeting system involved a monoclonal anti-PSA capture antibody-functionalized Fe 3 O 4 magnetic bead (mAb 1 -MB) and a polyclonal…

Fourier transform infrared spectroscopy (FTIR) is a ubiquitous spectroscopic technique. Spectral interpretation is a time-consuming process, but it yields important information about functional groups present in compounds and in complex substances. We develop a generalizable model via a machine learning (ML) algorithm using convolutional neural networks (CNNs) to identify the presence of functional groups in gas-phase FTIR spectra. The ML models reduce the amount of time required to analyze functional groups and facilitate interpretation of FTIR spectra. Through web scraping, we acquire intensity-frequency data from 8728 gas-phase organic molecules within the NIST spectral database and transform the data into spectral images. We successfully train models for 15 of the most common organic functional groups, which we then determine via identification from previously untrained spectra. These models serve to expand the application of FTIR measurements for facile analysis of organic samples. Our approach was done such that we have broad functional group models that infer in tandem to…

Elucidating the intrinsic relationship between diseases and lipid droplet (LD) polarity remains a great challenge owing to the lack of the research on multiple disease models. Until now, the visualization of abnormal LD polarity in models of inflammation and clinical cancer patient samples has not been achieved. To meet the urgent challenge, we facilely synthesized a robust LD-specific and polarity-sensitive fluorescent probe ( LD - TTP ), which consists of a triphenylamine segment as an electron-donor group (D) and a pyridinium as an electron-acceptor moiety (A), forming a typical D−π–A molecular configuration. Owing to the unique intramolecular charge transfer effect, LD-TTP exhibits high sensitivity to polarity change in the linear range from Δ f = 0.258 to 0.312, with over 278-fold fluorescence enhancement. Moreover, we revealed that LD-TTP possessed satisfactory ability for sensitively monitoring LD-polarity changes in living cells. Using LD-TTP, we first demonstrated the detection of LD-polarity changes in fatty liver tissues and inflammatory living mice via confocal laser…

Nuclear magnetic resonance (NMR) spectroscopy is a well-established method for analyzing protein structure, interaction, and dynamics at atomic resolution and in various sample states including solution state, solid state, and membranous environment. Thanks to rapid NMR methodology development, the past decade has witnessed a growing number of protein NMR studies in complex systems ranging from membrane mimetics to living cells, which pushes the research frontier further toward physiological environments and offers unique insights in elucidating protein functional mechanisms. In particular, in-cell NMR has become a method of choice for bridging the huge gap between structural biology and cell biology. Herein, we review the recent developments and applications of NMR methods for protein analysis in close-to-physiological environments, with special emphasis on in-cell protein structural determination and the analysis of protein dynamics, both difficult to be accessed by traditional methods.

Short of a vaccine, frequent and rapid testing, preferably at home, is the most effective strategy to contain the COVID-19 pandemic. Herein, we report on single-stage and two-stage molecular diagnostic tests that can be carried out with simple or no instrumentation. Our single-stage amplification is reverse transcription-loop mediated isothermal amplification (RT-LAMP) with custom-designed primers targeting the ORF1ab and the N gene regions of the virus genome. Our new two-stage amplification, dubbed Penn-RAMP, comprises recombinase isothermal amplification (RT-RPA) as its first stage and LAMP as its second stage. We compared various sample preparation strategies aimed at deactivating the virus while preserving its RNA and tested contrived and patient samples, consisting of nasopharyngeal swabs, oropharyngeal swabs, and saliva. Amplicons were detected either in real time with fluorescent intercalating dye or after amplification with the intercalating colorimetric dye LCV, which is insensitive to sample's PH. Our single RT-LAMP tests can be carried out instrumentation-free. To enable…

Raman spectral libraries specific to microplastics demonstrated improved spectral matching results when weathered plastics and a variety of particle colors and morphologies were included. Here, we explore if this is true for Fourier transform infrared (FTIR) spectroscopy as well. We present two novel databases specific to microplastics using attenuated total reflection (μATR-FTIR): (1) an FTIR library of plastic particles (FLOPP), containing 186 spectra from common plastic items, across 14 polymer types and (2) an FTIR library of plastic particles sourced from the environment (FLOPP-e), containing 195 spectra across 15 polymer types. Both libraries include particles from a variety of sources, morphologies, and colors. We demonstrate the applicability of these libraries for microplastics research by comparing spectral match results from two microplastic datasets. For this, we use different combinations of libraries including: commercially available reference libraries, an open-access polymer library, and FLOPP and FLOPP-e. Among tests, the greatest mean HQI result was achieved when…

Covid-19 pandemic outbreak is the reason of the current world health crisis. The development of effective antiviral compounds and vaccines requires detailed descriptive studies of SARS-CoV-2 proteins. The SARS-CoV-2 spike (S) protein mediates virion binding to the human cells through its interaction with the ACE2 cell surface receptor and is one of the prime immunization targets. A functional virion is composed of three S1 and three S2 subunits created by furin cleavage of the spike protein at R682, a polybasic cleavage site that differs from the SARS-CoV spike protein of 2002. By analysis of the protein produced in HEK293 cells, we observe that the spike is O-glycosylated on a threonine (T678) near the furin cleavage site occupied by core-1 and core-2 structures. In addition, we have identified eight additional O-glycopeptides on the spike glycoprotein and confirmed that the spike protein is heavily N-glycosylated. Our recently developed liquid chromatography-mass spectrometry methodology allowed us to identify LacdiNAc structural motifs on all occupied N-glycopeptides and…

Three-dimensional (3D)-printing techniques such as stereolithography (SLA) are currently gaining momentum for the production of miniaturized analytical devices and molds for soft lithography. However, most commercially available SLA resins inhibit polydimethylsiloxane (PDMS) curing, impeding reliable replication of the 3D-printed structures in this elastomeric material. Here, we report a systematic study, using 16 commercial resins, to identify a fast and straightforward treatment of 3D-printed structures and to support accurate PDMS replication using UV and/or thermal post-curing. In-depth analysis using Raman spectroscopy, nuclear magnetic resonance, and high-resolution mass spectrometry revealed that phosphine oxide-based photo-initiators, leaching out of the 3D-printed structures, are poisoning the Pt-based PDMS catalyst. Yet, upon UV and/or thermal treatments, photo-initiators were both eliminated and recombined into high molecular weight species that were sequestered in the molds.

Diabetes is one of the metabolic diseases marked by hyperglycemia and is often accompanied by the occurrence of some complications. As a biomarker of oxidative stress, hydrogen peroxide (H 2 O 2 ) has close association with the occurrence and development of diabetes and its complications. Unfortunately, there is no fluorescent probe reported for imaging H 2 O 2 in diabetic mice. Here, a novel near-infrared (NIR) fluorescent probe named QX-B was designed and synthesized to detect H 2 O 2 . For the probe, the quinolinium-xanthene dye is used as the fluorophore and borate ester is chosen as the response group. After the addition of H 2 O 2, a strong NIR fluorescence signal at 772 nm is observed. The probe not only shows high sensitivity with 10-fold enhancement but also displays excellent selectivity to H 2 O 2 over other possible interfering species. In the meantime, the possible response mechanism of QX-B toward H 2 O 2 was proposed and verified by the high-performance liquid chromatography (HPLC) experiment, mass spectra (MS) experiment, and density functional theory (DFT)…

The detection of microRNA (miRNA) in human serum has great significance for cancer prevention. Herein, a novel self-powered biosensing platform is developed, which effectively integrates an enzymatic biofuel cell (EBFC)-based self-powered biosensor with a matching capacitor for miRNA detection. A catalytic hairpin assembly and hybrid chain reaction are used to improve the analytical performance of EBFC. Furthermore, the matching capacitor is selected as an auxiliary signal amplifying device, and graphdiyne is applied as substrate material for EBFC. The results confirm that the developed method obviously increases the output current of EBFC, and the sensitivity can reach 2.75 μA/pM, which is 786% of pure EBFC. MiRNA can be detected in an expanded linear range of 0.1-100000 fM with a detection limit of 0.034 fM (S/N = 3). It can offer a selective and sensitive platform for nucleotide sequence detection with great potential in clinical diagnostics.

The development of a sensing platform with high sensitivity and specificity, especially programmability and universal applicability, for the detection of clinically relevant molecules is highly valuable for disease monitoring and confirmation but remains a challenge. Here, for the first time, we introduce the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system into an immobilization-free electrochemical biosensing platform for sensitively and specifically detecting the disease-related nucleic acids and small molecules. In this strategy, a modular rolling circle amplification (RCA) is designed to transform and amplify the target recognition event into the universal trigger DNA strand that is used as the trigger to activate the deoxyribonuclease activity of CRISPR/Cas12a for further signal amplification. The cleavage of the target-activated blocker probe allows the methylene blue-labeled reporter probes to be captured by the reduced graphene oxide-modified electrode, leading to an obviously increased electrochemical signal. We only need to simply tune the…

CRISPR-diagnostic assays have gained significant interest in the last few years. This interest has grown rapidly during the current COVID-19 pandemic, where CRISPR-diagnostics have been frontline contenders for rapid testing solutions. This surge in CRISPR-diagnostic research prompts the following question: what exactly are the achievable limits of detection and associated assay times enabled by the kinetics of enzymes such as Cas12 and Cas13? To explore this question, we here present a model based on Michaelis-Menten enzyme kinetics theory applied to CRISPR enzymes. We use the model to develop analytical solutions for reaction kinetics and develop back-of-the-envelope criteria to validate and check for consistency in reported enzyme kinetic parameters. We applied our analyses to all studies known to us, which report Michaelis-Menten-type kinetic data for CRISPR-associated enzymes. These studies include all subtypes of Cas12 and Cas13 and orthologs. We found all but one study clearly violate at least two of our three rules and therefore present data that violate basic physical…

Taking advantage of the excellent trans-cleavage activity, CRISPR-based diagnostics (CRISPR-Dx) has shown great promise in molecular diagnostics. However, the single-stranded DNA reporter of the current CRISPR-Dx suffers from poor stability and limited sensitivity, which make their application in complex biological environments difficult. Herein, we, for the first time, explore the trans-cleavage activity of CRISPR/Cas12a toward the substrate on gold nanoparticles and apply the new phenomenon to develop a spherical nucleic acid (SNA) reporter for stable and sensitive CRISPR-Dx biosensing. By anchoring the DNA substrate on gold nanoparticles, we discovered different trans-cleavage activities of different types of the Cas12a system (e.g., LbCas12a and AsCas12a) on a nanoparticle surface. The further study suggests that the trans-cleavage activity of LbCas12a on the nanoparticle surface is highly dependent on the density and length of DNA strands. Based on these interesting discoveries, we furthermore develop SNA reporter-based fluorescent CRISPR-Dx for stable and sensitive biosensing…

ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTDark-Field Microscopy: Recent Advances in Accurate Analysis and Emerging ApplicationsPeng Fei GaoPeng Fei GaoKey Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaMore by Peng Fei GaoView Biographyhttp://orcid.org/0000-0001-6878-6597, Gang LeiGang LeiKey Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, ChinaMore by Gang LeiView Biography, and Cheng Zhi Huang*Cheng Zhi HuangKey Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China*Email: [email protected]More by Cheng Zhi HuangView Biographyhttp://orcid.org/0000-0002-1260-5934Cite this: Anal. Chem. 2021, 93, 11, 4707–4726Publication Date (Web):February 23, 2021Publication History Published online23…

MicroRNA (miRNA) has emerged as one of the ideal target biomarker analytes for cancer detection because its abnormal expression is closely related to the occurrence of many cancers. In this work, we combined three-dimensional (3D) popcorn-like gold nanofilms as novel surface-enhanced Raman scattering (SERS)-electrochemistry active substrates with toehold-mediated strand displacement reactions (TSDRs) to construct a DNA molecular machine for SERS-electrochemistry dual-mode detection of miRNA. 3D popcorn-like spatial structures generated more active "hot spots" and thus enhanced the sensitivity of SERS and electrochemical signals. Besides, the TSDRs showed high sequence-dependence and high specificity. The addition of target miRNA will trigger the molecular machine to perform two TSDRs in the presence of signal DNA strands modified by R6G (R6G-DNA), thus achieving an enzyme-free amplification detection of miRNA with a low limit of detection of 0.12 fM (for the SERS method) and 2.2 fM (for the electrochemical method). This biosensor can also serve as a universally amplified and…

The global pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has revealed the urgent need for accurate, rapid, and affordable diagnostic tests for epidemic understanding and management by monitoring the population worldwide. Though current diagnostic methods including real-time polymerase chain reaction (RT-PCR) provide sensitive detection of SARS-CoV-2, they require relatively long processing time, equipped laboratory facilities, and highly skilled personnel. Laser-scribed graphene (LSG)-based biosensing platforms have gained enormous attention as miniaturized electrochemical systems, holding an enormous potential as point-of-care (POC) diagnostic tools. We describe here a miniaturized LSG-based electrochemical sensing scheme for coronavirus disease 2019 (COVID-19) diagnosis combined with three-dimensional (3D) gold nanostructures. This electrode was modified with the SARS-CoV-2 spike protein antibody following the proper surface modifications proved by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM)…

Alzheimer’s disease (AD) and Parkinson’s disease (PD) are chronic neurodegenerative diseases with high morbidity and mortality. Homocysteine (Hcy), cysteine (Cys), and glutathione (GSH) are closely related to AD and PD. However, the dynamics of Hcy, Cys, and GSH in the brain tissues and the potential pathogenesis between Cys/Hcy/GSH with AD and PD remain unclear. Herein, a novel fluorescent probe 1 with multiple binding sites was rationally designed and exploited for the direct quantification of serum total Hcy and Cys along with superior optical properties. Importantly, differentiation and simultaneity fluorescence imaging of Cys, Hcy, and GSH dynamics were achieved in living cells, tissues, and mouse models of AD and PD with this probe, providing direct evidences for the relationship between Hcy/Cys/GSH and AD/PD for the first time. In addition, pathogenesis studies demonstrated that elevated Hcy and Cys levels are closely related to imbalanced redox homeostasis, increased amyloid aggregates, and nerve cell cytotoxicity. These findings will greatly promote the understanding of the…

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global health emergency, and its gene mutation and evolution further posed uncertainty of epidemic risk. Herein, we reported a light-up CRISPR-Cas13 transcription amplification method, which enables the detection of SARS-CoV-2 and its mutated variants. Sequence specificity was ensured by both the ligation process and Cas13a/crRNA recognition, allowing us to identify viral RNA mutation. Light-up RNA aptamer allows sensitive output of amplification signals via target-activated ribonuclease activity of CRISPR-Cas13a. The RNA virus assay has been designed to detect coronavirus, SARS-CoV-2, Middle East respiratory syndrome (MERS), and SARS, as well as the influenza viruses such as, H1N1, H7N9, and H9N2. It was accommodated to sense as low as 82 copies of SARS-CoV-2. Particularly, it allowed us to strictly discriminate key mutation of the SARS-CoV-2 variant, D614G, which may induce higher epidemic and pathogenetic risk. The proposed RNA virus assays are promising for point-of-care monitoring of…

) in liquid chromatography (LC) among different laboratories, to provide supplementary evidence for the identity of compounds in high-resolution mass spectrometry (HRMS)-based suspect and nontarget screening investigations. In this study, a rigorously tested, flexible, and less system-dependent unified retention time index (RTI) approach for LC is presented, based on the calibration of the elution pattern. Two sets of 18 calibrants were selected for each of ESI+ and ESI-based on the maximum overlap with the retention times and chemical similarity indices from a total set of 2123 compounds. The resulting calibration set, with RTI set to range between 1 and 1000, was proposed as the most appropriate RTI system after rigorous evaluation, coordinated by the NORMAN network. The validation of the proposed RTI system was done externally on different instrumentation and LC conditions. The RTI can also be used to check the reproducibility and quality of LC conditions. Two quantitative structure-retention relationship (QSRR)-based models were built based on the developed RTI systems, which…

ECL is luminescence generated by electrochemical reactions and for this reason it possesses better spatio-temporal control and low background in comparison with photoluminescence or other optical methods that rely on external light illumination.1–3 In the last 20 years ECL has proved to be a versatile and powerful analytical technique in different fields, ranging from fundamental research to commercial clinical and biological applications.4,5 The main reason behind its success is that ECL offers remarkable advantages in comparison to other transduction methods: high sensitivity, an extremely wide dynamic range, very low background signal, good temporal and spatial control, and insensitivity to matrix effects.6,7 Thanks to its simplified optical setup, ECL has been implemented as a powerful imaging technique to visualize electrochemical objects and entities bringing important insight in the ECL mechanism generation. Here we aim to incorporate all the work done in the field of ECL imaging mainly in the last three years with a particular focus on ECL generation mechanisms and their…

Epilepsy is a chronic neurodegenerative disease that has seriously threatened human health. Accumulating evidence reveals that the pathological progression of epilepsy is closely related to peroxynitrite (ONOO – ). Unfortunately, understanding the physiological roles of ONOO – in epilepsy is still challenging due to the lack of powerful imaging probes for the determination of the level of fluctuations of ONOO – in the epileptic brain. Herein, a near-infrared (NIR) two-photon (TP) fluorescent probe [dicyanomethylene-4 H -pyran (DCM)–ONOO] is presented to trace ONOO – in living cells and in kainate (KA)-induced rat epilepsy models with satisfactory sensitivity and selectivity. The probe is composed of a NIR TP DCM fluorophore and a recognition moiety diphenylphosphinamide. The phosphoramide bond of the probe is interrupted after reacting with ONOO – for 10 min, and then, the released amino groups emit strong fluorescence due to the restoration of the intramolecular charge transfer process. The probe can effectively detect the changes of endogenous ONOO – with excellent temporal and…

Herein, an electrochemical (EC)–photoelectrochemical (PEC) dual-mode biosensor was constructed for cytokeratin 19 fragment 21-1 (CYFRA21-1) assay based on the dual-signaling electrochemical ratiometric strategy and “on–off–on” PEC method. The indium tin oxide (ITO) electrode was modified by 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA)@C 60 and gold nanoparticles (Au NPs), and the double-stranded DNA composed of thiol/methylene blue (MB)-labeled single-stranded DNA (ssDNA) (S0-MB) and antibody/ferrocene (Fc)-labeled ssDNA (Ab1-S1-Fc) was immobilized on the Au NPs/PTCDA@C 60 /ITO electrode via the Au–S bond between Au NPs and thiol of S0-MB. With the help of another antibody-labeled ssDNA (Ab2-S2), the presence of CYFRA21-1 triggered a typical antigen–antibody sandwich immune reaction (Ab1, CYFRA21-1, and Ab2) and proximity hybridization between Ab1-S1-Fc and Ab2-S2. This caused the release of Ab1-S1-Fc from the modified electrode and the change of S0-MB to a hairpin structure, resulting in a decrease (an increase) of the oxidation peak current of Fc (MB) and an increase of…

To elucidate the complex role of biological H 2 S and study the mitochondrial damage and some related diseases, effective methods for visualization of H 2 S in mitochondria and in vivo are urgently needed. In this contribution, a novel near-infrared mitochondria-targetable fluorescence probe MI-H 2 S for H 2 S detection was developed. MI-H 2 S shows rapid detection ability for H 2 S in pure aqueous solution and outputs a highly selective and sensitive fluorescence-on signal at 663 nm with a large Stokes shift of 141 nm. Bioimaging experiments revealed that the probe has good mitochondrial-targeting ability and high-contrast imaging ability for detecting H 2 S in living systems. The probe also showed great potential in the detection of H 2 S during inflammation. All of the results demonstrate that MI-H 2 S can be applied as an effective probe for the visualization and study of H 2 S in mitochondria and in vivo.

Non-targeted analysis (NTA) workflows using mass spectrometry are gaining popularity in many disciplines, but universally accepted reporting standards are nonexistent. Current guidance addresses limited elements of NTA reporting-most notably, identification confidence-and is insufficient to ensure scientific transparency and reproducibility given the complexity of these methods. This lack of reporting standards hinders researchers' development of thorough study protocols and reviewers' ability to efficiently assess grant and manuscript submissions. To overcome these challenges, we developed the NTA Study Reporting Tool (SRT), an easy-to-use, interdisciplinary framework for comprehensive NTA methods and results reporting. Eleven NTA practitioners reviewed eight published articles covering environmental, food, and health-based exposomic applications with the SRT. Overall, our analysis demonstrated that the SRT provides a valid structure to guide study design and manuscript writing, as well as to evaluate NTA reporting quality. Scores self-assigned by authors fell within the range of…

Rheumatoid arthritis (RA) is a common chronic autoimmune inflammatory disease, and its etiology is closely related to the overproduction of hypochlorous acid (HClO). However, early detection of RA using an activatable near-infrared-II (NIR-II, 1000–1700 nm) fluorescent probe remains challenging. Herein, we first report an “OFF–ON” NIR-II fluorescent probe named PTA (phenothiazine triphenylamine) for imaging HClO in deep-seated early RA. Electron-rich phenothiazine in the core of PTA was utilized as both an HClO-recognition moiety and a precursor of electron acceptors, displaying a typical donor–acceptor–donor structure with excellent NIR-II emission at 936/1237 nm once reacted with HClO. The probe PTA exhibited good water solubility, high photostability, and rapid response capability toward HClO within 30 s. Moreover, it was able to sensitively and specifically detect exogenous and endogenous HClO in living cells in both visible and NIR-II windows. Notably, PTA enabled the sensitive and rapid visualization of HClO generation in an inflammatory RA mouse model, showing a 4.3-fold…

A facile strategy for the electrochemical detection of human epidermal growth factor receptor 2 (HER2), a breast cancer biomarker, was presented via the fabrication of an antifouling sensing interface based on the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) and a biocompatible peptide hydrogel. The peptide hydrogel prepared from a designed short peptide of Phe-Glu-Lys-Phe functionalized with a fluorene methoxycarbonyl group (Fmoc-FEKF) enabled excellent activity preservation for the immobilized biomolecules, and its good hydrophilicity facilitated effective alleviation of nonspecific adsorption or biofouling, while the PEDOT film provided a highly stable and conducting substrate. The developed biosensor was highly sensitive and selective for HER2 detection, with a wide linear response range from 0.1 ng mL –1 to 1.0 μg mL –1 and a low limit of detection of 45 pg mL –1 . Moreover, the peptide hydrogel based biosensor was feasible to use for complex biological samples, and it was capable of detecting HER2 in human serum with clinically acceptable accuracy, manifesting a…

Respective detection of microplastics (MPs) and nanoplastics (NPs) is of great importance for their different environmental behaviors and toxicities. Using spherical polystyrene (PS) and poly(methyl methacrylate) (PMMA) plastics as models, the efficiency for sequential isolation of MPs and NPs by membrane filtration and cloud-point extraction was evaluated. After filtering through a glass membrane (1 μm pore size), over 90.7% of MPs were trapped on the membrane, whereas above 93.0% of NPs remained in the filtrate. The collected MPs together with the glass membrane were frozen in liquid nitrogen, ground, and suspended in water (1 mL) and subjected to pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS) determination. The NPs in the filtrate were concentrated by cloud-point extraction, heated at 190 °C to degrade the extractant, and then determined by Py-GC/MS. For MPs and NPs spiked in pure water, the method detection limits are in the range of 0.05-1.9 μg/L. The proposed method is applied to analyze four real water samples, with the detection of 1.6-7.6 μg/L PS MPs and 0.6 μg/L…

Surface-enhanced Raman spectroscopy (SERS) is a promising ultrasensitive analysis technology due to outstanding molecular fingerprint identification. However, the measured molecules generally need to be adsorbed on a SERS substrate, which makes it difficult to detect weakly adsorbed molecules, for example, the volatile organic compound (VOC) molecules. Herein, we developed a kind of a SERS detection method for weak adsorption molecules with Au@ZIF-8 core-shell nanoparticles (NPs). The well-uniformed single- and multicore-shell NPs can be synthesized controllably, and the shell thickness of the ZIF-8 was able to be precisely controlled (from 3 to 50 nm) to adjust the distance and electromagnetic fields between metal nanoparticles. After analyzing the chemical and physical characterization, Au@ZIF-8 core-shell NPs were employed to detect VOC gas by SERS. In contrast with multicore or thicker-shell nanoparticles, Au@ZIF-8 with a shell thickness of 3 nm could efficiently probe various VOC gas molecules, such as toluene, ethylbenzene, and chlorobenzene. Besides, we were capable of…

Mitophagy plays a crucial role in maintaining intracellular homeostasis through the removal of dysfunctional mitochondria and recycling their constituents in a lysosome-degradative pathway, which leads to microenvironmental changes within mitochondria, such as the pH, viscosity, and polarity. However, most of the mitochondrial fluorescence viscosity probes only rely on electrostatic attraction and readily leak out from the mitochondria during mitophagy with a decreased membrane potential, thus easily leading to an inaccurate detection of viscosity changes. In this work, we report a mitochondria-immobilized NIR-emissive aggregation-induced emission (AIE) probe CS-Py-BC, which allows for an off–on fluorescence response to viscosity, thus enabling the real-time monitoring viscosity variation during mitophagy. This system consists of a cyanostilbene skeleton as the AIE active core and viscosity-sensitive unit, a pyridinium cation for the mitochondria-targeting group, and a benzyl chloride subunit that induces mitochondrial immobilization. As the viscosity increased from 0.903 cP (0%…

Abstract There is an urgent need for ultra-rapid testing regimens to detect the SARS-CoV-2 [Severe Acute Respiratory Syndrome Coronavirus 2] virus infections in real-time within seconds to stop its spread. Current testing approaches for this RNA virus focus primarily on diagnosis by RT-qPCR, which is time-consuming, costly, often inaccurate and impractical for general population rollout due to the need for laboratory processing. The latency until the test result arrives with the patient has led to further virus spread. Furthermore, latest antigen rapid tests still require 15 to 30 min processing time and are challenging to handle. Despite increased PCR-test and antigen-test efforts the pandemic has entered the worldwide second stage. Herein, we applied a superfast reagent-free and non-destructive approach of attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy with subsequent chemometric analysis to the interrogation of virus-infected samples. Contrived samples with inactivated gamma-irradiated Covid-19 virus particles at levels down to 1582 copies/ml…

Herein we report on a deep-learning method for the removal of instrumental noise and unwanted spectral artifacts in Fourier transform infrared (FTIR) or Raman spectra, especially in automated applications in which a large number of spectra have to be acquired within limited time. Automated batch workflows allowing only a few seconds per measurement, without the possibility of manually optimizing measurement parameters, often result in challenging and heterogeneous datasets. A prominent example of this problem is the automated spectroscopic measurement of particles in environmental samples regarding their content of microplastic (MP) particles. Effective spectral identification is hampered by low signal-to-noise ratios and baseline artifacts as, again, spectral post-processing and analysis must be performed in automated measurements, without adjusting specific parameters for each spectrum. We demonstrate the application of a simple autoencoding neural net for reconstruction of complex spectral distortions, such as high levels of noise, baseline bending, interferences, or distorted…

Tumor-derived exosomal miRNAs may have important functions in the onset and progression of cancers and are potential biomarkers for early diagnosis and prognosis monitoring. Yet, simple, sensitive, and label-free detection of exosomal miRNAs remains challenging. Herein, an ultrasensitive, label-free, and stable field-effect transistor (FET) biosensor based on a polymer-sorted high-purity semiconducting carbon nanotube (CNT) film is reported to detect exosomal miRNA. Different from conventional CNT FETs, the CNT FET biosensors employed a floating gate structure using an ultrathin Y 2 O 3 as an insulating layer, and assembled Au nanoparticles (AuNPs) on Y 2 O 3 as linkers to anchor probe molecules. A thiolated oligonucleotide probe was immobilized on the AuNP surface of the sensing area, after which miRNA21 was detectable by monitoring the current change before and after hybridization between the immobilized DNA probe and target miRNA. This method achieved both high sensitivity (LOD: 0.87 aM) and high specificity. Furthermore, the FET biosensor was employed to test clinical plasma…

Nowadays, there is increased demand for wearable sensors for sweat glucose monitoring in order to facilitate diabetes management in a patient-friendly and noninvasive manner. This work describes a wearable glucose monitoring device in the form of an electrochemical ring (e-ring) fabricated by 3D printing. The 3D-printed e-ring consists of three carbon-based plastic electrodes (fabricated using a conductive filament) integrated at the inner side of a ring-shaped flexible plastic holder (fabricated using a nonconductive filament). The e-ring is modified with an electrodeposited gold film and is coupled to a miniature potentiostat directly addressable by a smartphone, offering the possibility for nonenzymatic amperometric self-testing of glucose levels in human sweat. Optical and electrochemical techniques are employed for the characterization of the e-ring. The device is resistant to mechanical bending and enables noninvasive glucose detection in sweat in the physiologically relevant concentration range of 12.5–400 μmol L –1 without interference from common electroactive metabolites.…

Biofouling caused by the accumulation of biomolecules on sensing surfaces is one of the major problems and challenges to realize the practical application of electrochemical biosensors, and an effective way to counter this problem is the construction of antifouling biosensors. Herein, an antifouling electrochemical biosensor was constructed based on electropolymerized polyaniline (PANI) nanowires and newly designed peptides for the detection of the COVID-19 N-gene. The inverted Y-shaped peptides were designed with excellent antifouling properties and two anchoring branches, and their antifouling performances against proteins and complex biological media were investigated using different approaches. Based on the biotin–streptavidin affinity system, biotin-labeled probes specific to the N-gene (nucleocapsid phosphoprotein) of COVID-19 were immobilized onto the peptide-coated PANI nanowires, forming a highly sensitive and antifouling electrochemical sensing interface for the detection of COVID-19 nucleic acid. The antifouling genosensor demonstrated a wide linear range (10 –14 to 10 –9…